Chromophore assisted laser inactivation of cellular proteins

Daniel G. Jay; F. S. Wang; H. Y. Chang; A. M. Sydor; J. C. Liao

doi:10.1117/12.274336

23 May 1997 Chromophore assisted laser inactivation of cellular proteins

Daniel G. Jay, F. S. Wang, H. Y. Chang, A. M. Sydor, J. C. Liao

Proceedings Volume 2983, Functional Imaging and Optical Manipulation of Living Cells; (1997) https://doi.org/10.1117/12.274336
Event: BiOS '97, Part of Photonics West, 1997, San Jose, CA, United States

Abstract

A molecular understanding of biology requires that we establish the in situ functions of the proteins in cellular processes. To address this, we developed chromophore- assisted laser inactivation (CALI) for probing the in vivo function of proteins. CALI inactivates specific proteins in living cells by using non-blocking antibodies conjugated with malachite green (MG) dye. MG absorbs 620 nm laser light (which is not absorbed by cells) to generate short lived free radicals with limited range of oxidative damage (15 angstroms) around the dye. This inactivates the bound protein without significantly affecting its neighbors. CALI has been applied to 40 proteins and achieved specific inactivation in almost all those tested. We have developed micro-CALI which uses a focused laser beam (10 micrometers ) to acutely inactivate specific proteins within cells. We have used this to address the molecular mechanisms of neuronal growth cone motility and has implicated a diversity of proteins (e.g. molecular motors, cytoskeletal, and signaling molecules) in discrete steps of growth cone motility. We hope that micro-CALI will be a useful research tool for addressing dynamic processes in biology and medicine.

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Daniel G. Jay, F. S. Wang, H. Y. Chang, A. M. Sydor, and J. C. Liao "Chromophore assisted laser inactivation of cellular proteins", Proc. SPIE 2983, Functional Imaging and Optical Manipulation of Living Cells, (23 May 1997); https://doi.org/10.1117/12.274336

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KEYWORDS

Proteins

Cell mechanics

Neurons

Diamond

In vivo imaging

Chromophores

In vitro testing

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